背景:钙电压门控通道辅助亚基α2/δ1(CACNA2D1),编码电压门控钙通道的基因,已被报道为几种癌症的癌基因。然而,其在结肠癌(CC)中的作用尚不清楚。本研究旨在探讨CACNA2D1的功能及其对CC微环境的影响。
方法:对200例接受根治性结肠切除术的CC患者的样本进行免疫组织化学(IHC)分析。在人CC细胞系HCT116和RKO中使用CACNA2D1siRNA进行敲除实验,和细胞增殖,循环,凋亡,然后对迁移进行了分析。将成纤维细胞系CCD-18Co与CC细胞系共培养,以确定CACNA2D1对成纤维细胞的影响以及CACNA2D1与癌症微环境之间的关系。使用微阵列分析来分析细胞的基因表达谱。
结果:IHC显示,高CACNA2D1表达是CC患者的独立不良预后因素,CACNA2D1表达与基质相关。CACNA2D1耗竭降低了细胞增殖和迁移;CACNA2D1敲除增加了处于亚G1期的细胞数量并诱导了细胞凋亡。CCD-18Co和HCT116或RKO细胞共培养表明,CACNA2D1通过成纤维细胞调节影响癌症微环境。此外,微阵列分析显示,p53信号通路和上皮-间质转化相关通路在CACNA2D1耗尽的HCT116细胞中增强.
结论:CACNA2D1通过调节成纤维细胞在CC的进展和微环境中发挥重要作用,可能作为疾病进展的生物标志物和CC的治疗靶标。
BACKGROUND: Calcium voltage-gated channel auxiliary subunit alpha 2/delta 1 (CACNA2D1), a gene encoding a voltage-gated calcium channel, has been reported as an oncogene in several cancers. However, its role in colon cancer (CC) remains unclear. This study aimed to investigate the function of CACNA2D1 and its effect on the microenvironment in CC.
METHODS: Immunohistochemistry (IHC) analysis was performed on samples collected from 200 patients with CC who underwent curative colectomy. Knockdown experiments were performed using CACNA2D1 siRNA in the human CC cell lines HCT116 and RKO, and cell proliferation, cycle, apoptosis, and migration were then analyzed. The fibroblast cell line CCD-18Co was co-cultured with CC cell lines to determine the effect of CACNA2D1 on fibroblasts and the relationship between CACNA2D1 and the cancer microenvironment. Gene expression profiles of cells were analyzed using microarray analysis.
RESULTS: IHC revealed that high CACNA2D1 expression was an independent poor prognostic factor in patients with CC and that CACNA2D1 expression and the stroma are correlated. CACNA2D1 depletion decreased cell proliferation and migration; CACNA2D1 knockdown increased the number of cells in the sub-G1 phase and induced apoptosis. CCD-18Co and HCT116 or RKO cell co-culture revealed that CACNA2D1 affects the cancer microenvironment via fibroblast regulation. Furthermore, microarray analysis showed that the p53 signaling pathway and epithelial-mesenchymal transition-associated pathways were enhanced in CACNA2D1-depleted HCT116 cells.
CONCLUSIONS: CACNA2D1 plays an important role in the progression and the microenvironment of CC by regulating fibroblasts and may act as a biomarker for disease progression and a therapeutic target for CC.